摘要
目的探讨胃癌成纤维细胞中透明质酸合成酶(HAS2)对胃癌细胞转移的影响。方法从手术切除的胃癌组织及癌旁正常组织中分别分离及培养胃成纤维细胞(CAFs)及正常成纤维细胞(NFs)。蛋白质免疫印迹(Western blot)及免疫荧光染色检测CAFs与NFs中α-平滑肌肌动蛋白(α-SMA)、波形蛋白(vimentin)、E-钙黏蛋白(E-cadherin)及HAS2的表达。利用siRNA下调CAFs中HAS2的表达,Western blot及免疫荧光染色检测CAFs中HAS2的表达,细胞迁移(Transwell)及划痕闭合实验检测胃癌细胞SGC7901的侵袭及迁移能力,Western blot检测SGC7901细胞上皮-间质转化相关蛋白E-cadherin、vimentin及N-钙黏蛋白(N-cadherin)的表达。结果α-SMA与HAS2在CAFs中高表达,vimentin在CAFs与NFs中均表达,而E-cadherin在CAFs与NFs中不表达。与si-NC组相比,si-HAS2组CAFs中HAS2表达降低,SGC7901细胞的侵袭及迁移能力降低,SGC7901细胞中上皮-间质转化相关蛋白E-cadherin表达升高,而Vimentin及N-cadherin表达降低。结论下调胃癌CAFs中HAS2的表达,可抑制胃癌细胞的侵袭、迁移及上皮-间质转化。
Objective To explore the effect of HAS2 in gastric cancer fibroblasts on the metastasis of gastric cancer cells.Methods CAFs and NFS were isolated and cultured from resected gastric cancer tissues and adjacent normal tissues,respectively.Western blot and immunofluorescence staining were used to detect the expression ofα-SMA,vimentin,E-cadherin and HAS2 in CAFs and NFs.siRNA down-regulated the expression of HAS2 in CAFs,the expression of HAS2 in CAFs was detected by Western blot and immunofluorescence staining,the invasion and migration ability of gastric cancer cell line SGC7901 was tested by Transwell and scratch closure test,the expression of epithelial-mesenchymal transition related proteins E-cadherin,vimentin and N-cadherin in SGC7901 cells was detected by Western blot.Resultsα-SMA and HAS2 were highly expressed in CAFs,vimentin was expressed in both CAFs and NFs,and E-cadherin was not expressed in CAFs and NFs.Compared with the si-NC group,the expression of HAS2 in CAFs was reduced,the invasion and migration ability of gastric cancer cells was reduced,the expression of E-cadherin in gastric cancer cells was increased,while the expression of vimentin and N-cadherin was decreased in the si-HAS2 group.Conclusion Down-regulating the expression of HAS2 in CAFs of gastric cancer could inhibit the invasion,migration and epithelial-mesenchymal transition of gastric cancer cells.
作者
唐忠斌
苏彧
刘伟
TANG Zhong-bin;SU Yu;LIU Wei(Department of Gastroenterology,the First People′s Hospital of Xiantao City,Affiliated to Changjiang University,Hubei Province 433000,China)
出处
《现代消化及介入诊疗》
2021年第4期446-450,共5页
Modern Interventional Diagnosis and Treatment in Gastroenterology
基金
湖北省卫生计生委科研项目(WJ2019Q017)。
